Protein expression and purification Insect cells Hi5

Generally it has been difficult to isolate high-quality RNA from yeast because of problems disrupting the cells. Use of enzymes to disrupt cell wall can alter gene expression profiles. Therefore, physical disruption can result in high quality RNA for all downstream processing. Use of DNAse and proteinase K will remove traces of DNA contamination and proteins respectively.

Generally it has been difficult to isolate high-quality RNA from yeast because of problems disrupting the cells. Use of enzymes to disrupt cell wall can alter gene expression profiles. Therefore, physical disruption can result in high quality RNA for all downstream processing. Use of DNAse and proteinase K will remove traces of DNA contamination and proteins respectively.

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

Get tips on using pCW-LIC to perform Protein expression and purification Bacteria - Escherichia coli Fbxo7

Get tips on using IMPACT™ Kit to perform Protein expression and purification Bacteria - Escherichia coli GmrSD

Get tips on using pGS-21a Vector to perform Protein expression and purification Bacteria - Escherichia coli p19

Get tips on using IEF and 2-D Protein Standards to perform Protein Ladder IEF and 2-D Standards

Get tips on using pET-21a(+) DNA to perform Protein expression and purification Bacteria - Escherichia coli Prefoldin (PFD)

Get tips on using pQE-80L vector to perform Protein expression and purification Bacteria - Escherichia coli IFABP-Aβ

Get tips on using pPIC9K Pichia Vector to perform Protein expression and purification Yeast - Pichia pastoris N-APP